Leishmania donovani: proteasome-mediated down-regulation of methionine adenosyltransferase

被引:10
作者
Perez-Pertejo, Yolanda [1 ]
Alvarez-Velilla, Raquel [1 ]
Garcia Estrada, Carlos [1 ]
Balana-Fouce, Rafael [1 ]
Reguera, Rosa M. [1 ]
机构
[1] Univ Leon, Dept Farmacol & Toxicol INTOXCAL, E-24071 Leon, Spain
关键词
methionine adenosyltransferase-overexpressing strain; post-translational regulation; Leishmania spp; S-adenosylmethionine; ubiquitin; proteasome; S-ADENOSYLMETHIONINE SYNTHETASE; TRYPANOSOMA-BRUCEI; 20S PROTEASOME; ORNITHINE-DECARBOXYLASE; PROTOZOAN PARASITE; GENOME SEQUENCE; 26S PROTEASOME; GENE; EXPRESSION; SUBUNIT;
D O I
10.1017/S0031182011000862
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
Methionine adenosyltransferase (MAT) is an important enzyme for metabolic processes, to the extent that its product, S-adenosylmethionine (AdoMet), plays a key role in trans-methylation, trans-sulphuration and polyamine synthesis. Previous studies have shown that a MAT-overexpressing strain of Leishmania donovani controls AdoMet production, keeping the intracellular AdoMet concentration at levels that are compatible with cell survival. This unexpected result, together with the fact that MAT activity and abundance changed with time in culture, suggests that different regulatory mechanisms acting beyond the post-transcriptional level are controlling this protein. In order to gain an insight into these mechanisms, several experiments were carried out to explain the MAT abundance during promastigote cell growth. Determination of MAT turnover in cycloheximide (CHX)-treated cultures resulted in a surprising 5-fold increase in MAT turnover compared to CHX-untreated cultures. This increase agrees with a stabilization of the MAT protein, whose integrity was maintained during culture. The presence of proteasome inhibitors, namely MG-132, MG-115, epoxomycin and lactacystin in the culture medium prevented MAT degradation in both MAT-overexpressing and 'mock-transfected' leishmanial strains. The role of the ubiquitin (Ub) pathway in MAT down-regulation was supported using immunoprecipitation experiments. Immunoprecipitated MAT cross-reacted with anti-Ub antibodies, which provides evidence of a proteasome-mediated down-regulation of the leishmanial MAT abundance.
引用
收藏
页码:1082 / 1092
页数:11
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